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Title: Latest methods of fluorescence-based protein crystal identification

Fluorescence, whether intrinsic or by using trace fluorescent labeling, can be a powerful aid in macromolecule crystallization. Its use in screening for crystals is discussed here. Successful protein crystallization screening experiments are dependent upon the experimenter being able to identify positive outcomes. The introduction of fluorescence techniques has brought a powerful and versatile tool to the aid of the crystal grower. Trace fluorescent labeling, in which a fluorescent probe is covalently bound to a subpopulation (<0.5%) of the protein, enables the use of visible fluorescence. Alternatively, one can avoid covalent modification and use UV fluorescence, exploiting the intrinsic fluorescent amino acids present in most proteins. By the use of these techniques, crystals that had previously been obscured in the crystallization drop can readily be identified and distinguished from amorphous precipitate or salt crystals. Additionally, lead conditions that may not have been obvious as such under white-light illumination can be identified. In all cases review of the screening plate is considerably accelerated, as the eye can quickly note objects of increased intensity.
Authors:
;  [1] ;  [2] ;  [3]
  1. University of Hamburg, c/o DESY Building 22a, Notkestrasse 85, 22607 Hamburg (Germany)
  2. iXpressGenes Inc., 601 Genome Way, Huntsville, AL 35806 (United States)
  3. (Germany)
Publication Date:
OSTI Identifier:
22375712
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta crystallographica. Section F, Structural biology communications; Journal Volume: 71; Journal Issue: Pt 2; Other Information: PMCID: PMC4321462; PMID: 25664782; PUBLISHER-ID: en5560; PUBLISHER-ID: S2053230X15000114; OAI: oai:pubmedcentral.nih.gov:4321462; Copyright (c) Meyer et al. 2015; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALS; FLUORESCENCE; ILLUMINANCE; MODIFICATIONS; PLATES; PROBES; PROTEINS; REVIEWS; SALTS; SCREENING; VISIBLE RADIATION